NDLI: Radiometal labeling of recombinant proteins by a genetically engineered minimal chelation site: technetium-99m coordination by single-chain Fv antibody fusion proteins through a C-terminal cysteinyl peptide.

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Radiometal labeling of recombinant proteins by a genetically engineered minimal chelation site: technetium-99m coordination by single-chain Fv antibody fusion proteins through a C-terminal cysteinyl peptide.

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Radiometal labeling of recombinant proteins by a genetically engineered minimal chelation site: technetium-99m coordination by single-chain Fv antibody fusion proteins through a C-terminal cysteinyl peptide.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Jamar, F. Heelan, B. T. Peters, A. M. George, A. J. Adams, G. P. Oppermann, H. Mccartney, J. E. Tai, M. S. Weiner, L. M. Houston, L. L.
Description	Author Affiliation: George AJ ( Department of Immunology, Royal Postgraduate Medical School, Hammersmith Hospital, London, United Kingdom.);
Abstract	We describe a method to facilitate radioimaging with technetium-99m (99mTc) by genetic incorporation of a 99mTc chelation site in recombinant single-chain Fv (sFv) antibody proteins. This method relies on fusion of the sFv C terminus with a Gly4Cys peptide that specifically coordinates 99mTc. By using analogues of the 26-10 anti-digoxin sFv as our primary model, we find that addition of the chelate peptide, to form 26-10-1 sFv', does not alter the antigen-binding affinity of sFv. We have demonstrated nearly quantitative chelation of 0.5-50 mCi of 99mTc per mg of 26-10-1 sFv' (1 Ci = 37 GBq). These 99mTc-labeled sFv' complexes are highly stable to challenge with saline buffers, plasma, or diethylenetriaminepentaacetic acid. We find that the 99mTc-labeled 741F8-1 sFv', specific for the c-erbB-2 tumor-associated antigen, is effective in imaging human ovarian carcinoma in a scid mouse tumor xenograft model. This fusion chelate methodology should be applicable to diagnostic imaging with 99mTc and radioimmunotherapy with 186Re or 188Re, and its use could extend beyond the sFv' to other engineered antibodies, recombinant proteins, and synthetic peptides.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	18
Volume Number	92
Language	English
Publisher	National Academy of Sciences
Publisher Date	1995-10-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Cysteine Chemistry Immunoglobulin Fragments Peptides Recombinant Fusion Proteins Technetium Amino Acid Sequence Animals Chelating Agents Cloning, Molecular Genetics Kinetics Mice Mice, SCID Molecular Sequence Data Neoplasms, Experimental Protein Folding Radionuclide Imaging Pharmacokinetics Tissue Distribution Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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